Cell Cycle Assay Kit (Red)

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Afmeting	Eenheidsprijs	België	Japan*	Hoeveelheid
1KIT	€276.00	1	7

*Stock beschikbaar uit voorraad in België leverbaar in 1 tot 3 dagen
*stock beschikbaar uit voorraad in Japan leverbaar in 1 tot 2 weken (met uitzondering van gereguleerde producten en zendingen met droog ijs)

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Description

This product is supplied as a two separate components, a DNA-specific dye (7-AAD) used to stain intracellular DNA and a buffer that allows for staining of cells with or without prior fixation. The resultant stained cells can be analyzed via flow cytometry (488/561nm laser, 630nm-700nm filter) to visualize the proportion of cells in the G0/G1, S, and G2/M stages of the cell cycle, determined by the strength of DNA staining. Enough for 100 tests.

Contents

• 7-AAD: 1 vial
• Cell Staining Buffer: 1 vial

*Dispose of reagents according to your institutional regulations.

Directions for Use

1. Dissolve 7-AAD in 100 µL DMSO before first use. Aliquot and store at -20°C
2. For each sample to be analyzed, combine 200 µL Buffer with 1 µL 7-AAD.
3. Trypsinize and centrifuge cells. Or, if using ethanol-fixed cells, wash once with PBS and centrifuge. Dispose of supernatant.
4. To each sample of 2.0 x 10⁴ – 1.0 x 10⁵ cells, add 200 µL of the mixture from step 2 and allow to incubate at room temperature for 30 minutes*.
5. Analyze on a flow cytometer using a laser in the range of 488nm-590nm and a filter with a wavelength in the range of 613nm-700nm.

*Optimal incubation varies between cell lines; optimization may be required.

Precautions

Results may vary when using non-fixed suspension cells. Extra care must be taken not to accidentally lyse these cells during pipetting steps, and cells should be run as quickly after staining as possible.

Storage

Store staining buffer at 4°C. Over time, a white crystalline precipitate may form. Formation of this precipitate has no effect on assay results, however it must be redissolved by vortexing at room temperature before further use. Store both original non-dissolved cell stain and DMSO-dissolved cell stain at -20°C.